Construction of prokaryotic expression vector for Ag85A-HA2 fusion gene and studies on the immunity efficacy of fusion protein against influenza a virus

Shao, J.-J.; Yang, J.; Dai, J.; Meng, J.-J.; Pei, D.-C.; Li, H.; Pan, X.; Li, W.-Y.

Sichuan da Xue Xue Bao. Yi Xue Ban 45(4): 547-551

2014


ISSN/ISBN: 1672-173X
PMID: 25286673
Accession: 052313719

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Abstract
To construct Ag85A-HA2 prokaryotic expression vector, express the fusion protein and study the immunity efficacy of fusion protein against influenza A virus. Ag85A-HA2 prokaryotic expression vector was constructed and induced with IPTG. The fusion protein was identified by SDS-PAGE and purified with His-Tag affinity chromatography. The BALB/c mice were immunized with fusion protein. Then the pathological section, lung index, lung inhibitory rate and death-protection rate were tested to evaluate the immunity efficacy of fusion protein. pET-32a(+)/Ag85A-HA2 prokaryotic expression vector was constructed successfully. And SDS-PAGE indicated that fusion protein was expressed correctly with a molecular mass of 70 x 10(3). The lung index and death-protection rate in experimental group were 39.30% and 80%, higher than that of control group. The pathological section also demonstrated that Ag85A-HA2 fusion protein had a protective effect on murine lungs. Ag85A-HA2 prokaryotic expression vector was successfully constructed, inducible expression and the fusion protein had an immunity efficacy against influenza A virus in animal experiment.